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268 Chapter 6: Breast Cancer Tumor cell clonogenic assay The mononuclear cell fraction was placed in medium ready for plating. In some cases where multiple PBPC specimens were obtained, the specimens were pooled and processed as a single specimen. A total of 5.0XI0 5 to 1.0X10 6 mononuclear cells per mL were plated in triplicate in 35 mm 2 grid-bottomed petri dishes (Nunc, Naperville, IL) in a soft agar-based medium consisting of final concentration of 15% Iscove's modified Dulbecco's medium (IMDM) (Sigma), 10% fetal bovine serum (Sigma), 0.3% agar solution (Sigma), 2 pg/mL human recombinant epidermal growth factor (Collaborative Research, Bedford, MA), and 10 pg/mL human recombinant insulin-like growth factor I (Collaborative Research). Negative control plates consisted of medium without supplemental growth factors and PBPC specimens from patients with nonepithelial tumors (e.g., normal donors, neurob lastoma, etc.). All plates were incubated in a humidified chamber at 37°C with 7.5% C0 2 for 10-14 days. An inverted phase contrast microscope was used to count tumor colonies (>25 cells). Tumor colony identification To definitively distinguish tumor colonies from normal hematopoietic colonies, agar plates were floated onto 2- X 3-inch microscope slides and air-dried at 4°C. Initially, in situ tumor colony verification was performed using immunofluorescence staining with FITC-labeled anti-cytokeratin mAb SB-3 (Caltag, San Francisco, CA) (Fig. 1). In certain cases, granulocyte-macrophage colony-forming unit (CFU-GM) colony verification was performed using immunofluorescence staining with rhodamine-labeled anti-CD 11 mAb (Dako, Carpenteria, CA). Later, an immunocytochemical method was developed to verify and count the number of tumor colonies. Slides were placed into cold storage at -70°C. The patient specimens were then removed from storage and tested for immunoreactivity. Immunostaining was accomplished using the alkaline phosphatase-based technique. Slides were baked in an oven, fixed in a paraformaldehyde solution at 4°C for 30 minutes, and washed thoroughly in PBS with 1% triton (Gibco). After fixation, slides were incubated with agarase followed by anti-breast carcinoma antibodies for 45 minutes at room temperature. Biotek immunoreagents for alkaline phosphatase staining were performed as per protocol. The specimen was then incubated with Ehrlich's hematoxylin counterstain and coverslipped with mounting medium. The number of tumor colonies per plate was then recorded as identified by the above methods. PBPC specimens were excluded from analysis if the cell viability was
Moss et al. 269 0 15 70 143 200 400 600 800 900 Months Post -ASCT Figure 1. Survival curve comparing the disease-free survival of patients with an ICC-positive marrow to those with a negative sample. RESULTS Paired marrow and stem cell samples were available from 246 stage IV patients. All samples were evaluated using ICC, and 130 PBPC and 66 bone marrow samples were analyzed using the clonogenic assay. The marrow was positive in 43 instances (17.5%), and the PBPC was positive in 40 cases (16.2%). In 15 patients, the marrow and PBPC both had breast cancer cells detected. The marrow and PBPC were negative for tumor cells in 178 women (see Table 1). Clinical follow-up was available for 246 patients. The overall DFS was 15% for all patients at a median follow-up of 30 months. The median DFS was significantly reduced for patients with ICC-positive marrow (146 days) when compared with patients who were ICC-negative (458 days; P=0.0001; see Fig. 1). All 15 patients whose marrow and PBPC samples were positive for tumor relapsed (median DFS 286 days vs. 423 days; P=0.015). The patient group with the Table 1. The incidence of breast cancer cells in marrow and PBPC collections Specimen type ICC positive ICC negative Marrow 43 203 PBPC 40 246 Marrow and PBPC* 15 178 *53 patients had only one specimen for breast cancer cells (either the marrow or the PBPC).
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AUTOLOGOUS BLOOD AND MARROW TRANSPL
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AUTOLOGOUS BLOOD AND MARROW TRANSPL
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ACKNOWLEDGMENTS We wish to thank Wi
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THE VAN BEKKUM STEM CELL AWARD Prof
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xii Table of Contents CHAPTER 2: AL
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xiv Table of Contents Late Non-Rela
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xvi Table of Contents Pretreatment
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xviii Table of Contents CHAPTER 7:
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XX Table of Contents Long-Term Obse
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xxii Table of Contents CHAPTER 13:
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0-9 LIST OF ABBREVIATIONS 2CDA 2-ch
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List of Abbreviations xxvii EFS eve
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List of Abbreviations MOPP mechlore
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VOD veno-occlusive disease VPC viru
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4HC-Purged Autologous Stem Cell Tra
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1.0-1 + 0.2 Miller et al. 5 Event F
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Miller et al. Adjusted probability
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Who Benefits From Autograft in AML
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Burnett 11 which to set the transpl
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Table 1. Outcome after relapse in M
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Immunotherapy in AML: No Positive E
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Simonsson et al. inhibit IL-2 induc
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Autologous Bone Marrow Transplantat
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Ball et al. 21 CD15) and AML-2-23 (
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Ball et al. 23 The ex vivo treatmen
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Ball et al. 25 Table 2. Factors inv
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Ball et al. Overall Survival for Pa
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Ball et al. Overall Survival for Pa
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Ball et al. 31 Overall Survival for
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Ball et al. Table 4. Actuarial over
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Ball et al. myeloid leukemia. The G
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Ball et al. 1993. 38. Mehta J, Powl
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Can Autologous Stem Cell Transplant
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De Witte et al. 41 novo AML. Auer r
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De Witte et al. hematopoietic engra
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De Witte et al. 23. Fenaux P, Laï
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Stein et al. and fever. No patient
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Stein et al. after doses 1, 3, and
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Stein et al. early after the transp
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autologous stem-cell rescue. / Clin
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Bone Marrow Transplantation for Acu
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Table 2. Autologous BMT in ALL in f
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Rowe 61 or adult patients with acut
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Hoelzer and Gôkbuget minitransplan
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Hoelzer and Gdkbuget 65 blood, earl
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Table 3. Risk factors for the defin
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Hoelzer and Gôkbuget treatment opt
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Hoeker and Gökbuget tical sibling
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Sierra et al. INTRODUCTION Two-thir
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Sierra et al. Table 2. Adverse char
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1,0 0,0 J Sierra et al. 0 20 40 60
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Sierra et al. Months Figure 4 < 30,
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Sierra et al. No adverse factors (n
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1994. Sierra et al. 83 10. Canals C
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Martin, Atta, and Hoelzer 85 adult
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Martin, Atta, and Hoelzer 87 Single
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100. Martin, Atta, and Hoelzer 89 P
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Martin, Atta, and Hoelzer 91 chromo
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Autologous vs. Unrelated Allogeneic
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Boyer and Yeager 95 outcomes after
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Boyer and Y eager 97 Table 2. Hypot
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Boyer and Y eager SUMMARY Most of t
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CHAPTER 3 CML
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Autograft Followed by Allograft Wit
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106 Chapter 3: CML (one low-grade a
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108 Chapter 3: CML DISCUSSION Myelo
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110 Chapter 3: CML Philadelphia-neg
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The Case for Manipulation of CML Au
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Toward a Cure by Drug Treatment? Th
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116 Chapter 3 :CML Table 1. Prolong
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118 Chapter 3: CML Table 3. German
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120 Chapter 3: CML Table 5. Normal
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122 Chapter 3: CML Table 8. Surviva
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124 Chapter 3: CML 88:3118-3122, 19
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126 Chapter 3: CML RK, Klingemann H
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The Tyrphostin AG957 Inhibits the I
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130 Chapter 3: CML mobilization. Al
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132 Chapter 3: CML A B 100 n 0 1 5
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134 Chapter 3: CML The in vitro sel
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136 Chapter 3: CML detecting leukem
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Progressive Hodglcin's Lymphoma Fol
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Reece et al. survival is observed i
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0.2 -I 0.0 0 Reece et al. 143 ii i
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Reece et al. Table 3. Causes of non
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Reece et al. carmustine, etoposide
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CD34 + Selection of Hematopoietic B
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Lazarus et al. 151 significantly de
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Lazarus et al. 153 (Sigma, St Louis
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Lazarus et al. 155 Table 2. Effect
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Lazarus et al. 157 difference. Furt
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Lazarus et al. 159 cells after myel
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The European CUP/UP Trial: A Prelim
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Table 1. Patient characteristics at
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Schouten et al. licular non-Hodgkin
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Sweetenham et al. vs. autologous pe
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Table 1. Patient characteristics, g
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Table 2. Patient characteristics, g
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Table 3. Sites of relapse, group A
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Table 4. Patterns of relapse, group
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A 100 80 %OS 60 40 20 B 100 %OS | S
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Sweetenham et al. new sites, and fi
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Sweetenham et al. Treatment options
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Molecular Remission: A Worthwhile A
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Early or Late Autotransplant in Hig
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Schenkein et al. 187 PATIENTS AND M
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Schenkein et al. 189 Toxicity Toxic
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Schenkein et al. 191 16. Glick JH,
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Santini et al. 193 to evaluate the
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Table 1. Continued Santini et al. 1
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Santini et al. 197 cytosine arabino
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1 0 0 1 90- 80- 70- (0 u. 60' o so-
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Hodgkin's lymphoma. N EnglJ Med 333
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CHAPTER 5 MYELOMA
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206 Chapter 5: Myeloma independent
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208 Chapter 5: Myeloma disease (MRD
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210 Chapter 5: Myeloma Scandinavia,
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212 Chapter 5: Myeloma (0 n o B co
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214 Chapter 5: Myeloma B cq d ? o c
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216 Chapter 5: Myeloma patients wit
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Page 256 and 257: Autologous Transplantation in Multi
Page 258 and 259: 224 Chapter 5: Myeloma 0 52 44 U §
Page 260 and 261: 226 Chapter 5: Myeloma BM ^ (CD34±
Page 262 and 263: 228 Chapter 5: Myeloma IFM 94 : OS
Page 264 and 265: 230 Chapter 5: Myeloma two groups a
Page 266 and 267: 232 Chapter 5: Myeloma increase bot
Page 268 and 269: 234 Chapter 5: Myeloma REFERENCES 1
Page 270 and 271: 236 Chapter 5: Myeloma Intensified
Page 272 and 273: 238 Chapter 5: Myeloma of CD34 + ce
Page 274 and 275: Chapter 5: Myeloma Table 2. Descrip
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Page 278 and 279: 244 Chapter 5: Myeloma ated with sh
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Page 283 and 284: Kvalheim et al. 249 Table 1. Immuno
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Page 287 and 288: Kvalheim et al. 253 DISCUSSION In t
Page 289 and 290: Kvalheim et al. 255 Bastert G: Micr
Page 291 and 292: Hood et al. 257 In an effort to inc
Page 293 and 294: #CD34 + cells in blood = Hood et al
Page 295 and 296: Hood et al. 261 Table 3. Frequency
Page 297 and 298: Hood et al. 263 REFERENCES 1. Rill
Page 299 and 300: The Clinical Significance of Breast
Page 301: Moss et al. tests, and bilateral po
Page 305 and 306: Moss et al. 271 0 3 6 12 18 24 30 3
Page 307 and 308: Moss et al. 273 8. Passos-Coelho J,
Page 309 and 310: Autotransplantation for Men With Br
Page 311 and 312: McCarthy et al. 277 radiation, two
Page 313 and 314: High Dose Sequential Chemotherapy W
Page 315 and 316: Viens et al. 281 3 g/m 2 and doxoru
Page 317 and 318: Table 1. Tumor characteristics Exte
Page 319 and 320: Table 4. Response Viens et al. 285
Page 321 and 322: Viens et al. 287 Table 5. Pathologi
Page 323 and 324: Viens et al. 289 apy for inoperable
Page 325 and 326: Gliick et al. 291 INTRODUCTION The
Page 327 and 328: Gluck et al. 293 RESULTS Patient de
Page 329 and 330: Glück et al. 295 Table 3. Response
Page 331 and 332: Gluck et al. 297 Overall Survival O
Page 333 and 334: Gluck et al. 299 Sudbury patients w
Page 335 and 336: Gluck et al. 301 anthracycline or t
Page 337 and 338: Repetitive High-Dose Therapy With P
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Page 345 and 346: Table 4. Hematopoietic recovery fol
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Table 5. Results of Isolex 300i CD3
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Prince et al. 321 Figure 5, continu
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Prince et al. 323 months after tran
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Prince et al. 325 excessive to be u
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Prince et al. 327 eight patients ha
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Prince et al. 329 marrow transplant
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Prince et al. intensive chemotherap
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-Q CO .Q O 1.0 0.9 0.8 0.7 0.6 0.5
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.Q CD .Q O 1.0 0.9 0.8 0.7 0.6 0.5
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oba Q. 1.0 0.9 Dicke et al. Stage I
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Vahdat with peripheral blood progen
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Evaluation of Prognostic Factors fo
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Fields et al. 343 PATIENTS AND METH
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Table 1. Chemotherapy regimens. Fie
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Fields et al. Stage II; 4-9 Pos LN
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U U Fields et al.
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Fields et al. 351 between 1 and 2 y
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European Trends and the EBMT Databa
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25% % 22% / 17% \ Rosti et al. 355
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Rosti et al. 357 The new Italian St
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Rosti et al. 6. Haas R, Schmid H, H
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Bewick et al. 363 INTRODUCTION The
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Bewick et al. 365 Blood samples obt
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Bewick et al. 367 HDCT with ABSC su
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Bewick et al. Progression Free Surv
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Bewick et al. pression in MBC, i.e.
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Bewick et al. 14. Kandl HL, Seymour
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CHAPTER 7 OTHER SOLID TUMORS
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378 Chapter 7: Solid Tumors INTRODU
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380 Chapter 7: Solid Tumors Prepara
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382 Chapter 7: Solid Tumors Surviva
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384 Chapter 7: Solid Tumors months
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386 Chapter 7: Solid Tumors have be
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High-Dose Chemotherapy in the Manag
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390 Chapter 7: Solid Tumors followe
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392 Chapter 7: Solid Tumors Table 1
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394 Chapter 7: Solid Tumors (mucosi
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Three-Fold Intensification by Seque
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High-Dose Therapy for Small Cell Lu
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404 Chapter 7: Solid Tumors chemoth
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406 Chapter 7: Solid Tumors SCLC ha
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408 Chapter 7: Solid Tumors relapse
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410 Chapter 7: Solid Tumors 69:585-
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412 Chapter 7: Solid Tumors ogous b
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414 Chapter 7: Solid Tumors 64. Bru
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416 Chapter 7: Solid Tumors Prignot
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418 Chapter 7: Solid Tumors CO > CO
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420 Chapter 7: Solid Tumors TANDEM
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Multiple Courses of Cyclophosphamid
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424 Chapter 7: Solid Tumors and the
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426 Chapter 7: Solid Tumors Thiotep
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430 Chapter 7: Solid Tumors E 100 0
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432 Chapter 7: Solid Tumors course
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434 Chapter 7: Solid Tumors boplati
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436 Chapter 7: Solid Tumors INTRODU
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438 - Chapter 7: Solid Tumors Table
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440 Chapter 7: Solid Tumors seeded
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442 Chapter 7: Solid Tumors DISCUSS
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444 Chapter 7: Solid Tumors 10. Di
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446 Chapter 7: Solid Tumors plasma
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Autologous Stem Cell Transplantatio
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Keystone 451 hypertension, anemia,
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Fassas et al. 453 INTRODUCTION Mult
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Fassas et al. Table 2. Stem cell mo
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Table 3. Toxicity after stem cell i
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Fassas et al. Table 5. Trial 1: cha
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Fassas et al. MS PROGRESSION FREE S
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Fassas et al. ic or autologous bone
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1 2 1 6 EAE. Burt et al. 465 Becaus
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Burt et al. Since cyclophosphamide
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Burt et al. 469 toxicity study, we
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Burt et dl. All 23. Mor F, Cohen IR
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Hasler, Gratwohl, and Tyndall an ev
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Hasler, Gratwohl, and Tyndall 475 B
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Kuis, Wulffraat, and Sanders 477 st
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Kuis, Wulffraat, and Sanders neousl
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CHAPTER 9 LONG-TERM EFFECTS
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484 Chapter 9: Long-Term Effects po
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486 Chapter 9: Long-Term Effects Ta
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488 Chapter 9: Long-Term Effects ml
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490 Chapter 9: Long-Term Effects 4.
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492 Chapter 9: Long-Term Effects us
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494 Chapter 9: Long-Term Effects AB
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498 Chapter 9: Long-Term Effects En
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500 Chapter 9: Long-Term Effects Rl
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502 Chapter 9: Long-Term Effects RE
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504 Chapter 9: Long-Term Effects A,
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Long-Term Follow-Up of Autologous T
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CHAPTER 10 GRAFT MANIPULATION
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514 Chapter 10: Graft Manipulation
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Stem Cell Reinfusion Over Two Conse
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Final Report of the First Prospecti
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Canine Hematopoietic Stem Allograft
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Sandmaier et al. 603 The resultant
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Sandmaier et al. 605 synthesis path
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Sandmaier et al. 607 interactions o
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Sandmaier et al. 1991. 2. Burchenal
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87:3508-3513,1996. Sandmaier et al.
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Giralt, Khouri, and Champlin Table
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Giralt, Khouri, and Champlin seven
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1.0 3 0.2 Giralt, Khouri, and Champ
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CHAPTER 12 GENE THERAPY
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622 Chapter 12: Gene Therapy INTROD
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624 Chapter 12: Gene Therapy RESULT
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626 Chapter 12: Gene Therapy envisi
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628 Chapter 12: Gene Therapy cer in
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A Vaccine of Melanoma Peptide Loade
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B43(anti-CD 19)-Gen i stein Immunot
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Targeting Immunotherapy for the Tre
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McGuinness andArceci 637 modified m
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McGuinness andArceci 639 leukemic c
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McGuinness and Arced 641 in G418. T
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McGuinness andArceci 643 Figure 2
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mRNA for hB7-1 (1491 bp) mRNA for C
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McGuinness and Arced CD80PE C080PE
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McGuinness andArceci 649 8. Robert
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McGuinness andArceci immune respons
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McGuinness andArceci 75. Jubinsky F
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Slavin et al. 655 up suggests, as p
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Slavin et al. 657 was a phase lib c
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Slavin et al. presented, appears to
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Antitumor Immunotherapy in Autologo
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Klingemann et al. 663 Table 3. Meth
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CHAPTER 14 RADIOIMMUNOTHERAPY
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668 Chapter 14: Radioimmunotherapy
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Selection of Radioisotopes, Chelate
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Radioimmunotherapy of Common Epithe
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CHAPTER 15 NEW AVENUES
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698 Chapter 15: New Avenues Table 1
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700 Chapter 15: New Avenues An alte
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702 Chapter 15: New Avenues who can
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704 Chapter 15: New Avenues myeloid
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706 Chapter 15: New Avenues 48. Gir
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Emerging Viral Infections in Blood
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710 Chapter 15: New Avenues influen
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712 Chapter 15: New Avenues Among a
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714 Chapter 15: New Avenues physica
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716 Chapter 15: New Avenues respira
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Minimal Therapy Models of Transplan
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CHAPTER 16 SUMMARIES
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724 Chapter 16: Summaries although
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726 Chapter 16: Summaries use of po
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728 Chapter 16: Summaries Table 1.
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736 Chapter 16: Summaries IL-15. Th
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738 Chapter 16: Summaries Patterns
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740 List of Participants Thomas L.
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742 List of Participants Sergio A.
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744 List of Participants Hans Marti
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746 List of Participants David P. S
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748 List of Participants AUTHOR IND
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750 Author Index Hurd, D.D. 483 Kui
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752 Author Index Stiff, Patrick J.
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754 Key Word Index Hematologic mali
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ISBN 1-891524-04-6
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